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Stage Gids EPB 2020 research group: Evolutionary & Population Biology Research Projects The role of ants as pollinators and seed dispersers in the rare Thesium humifusum Supervisors: Gerard Oostermeijer & Sheila Luijten (Science4Nature) Contact: [email protected] or [email protected] Project outline Even though ants are very common members of nearly every biological community, they are remarkably scarce as pollinators, in particular in temperate regions. Even though they regularly visit flowers of various species, ants are generally considered poor pollinators because of their small size, low action radius and frequent grooming. Recently, the fact that ants secrete antibiotic compounds has also been associated with their limited role as pollinators. Nevertheless, specialized pollination relations occur between orchids and ants, and in harsh systems that favour low growth forms, ant pollination also occurs, albeit rarely. Ant dispersal (myrmecochory) is much more frequent in our flora, and many species have seeds bearing food bodies, or elaiosomes, that cause ants to carry them (in)to their nest. Because ants are small, and have a limited foraging range, the function of this dispersal is not so much attaining a distance from the mother plant, but more about being deposited in a safe site for germination and survival (directional dispersal). We study the demography and genetics of the extremely rare plant Thesium humifusum (Santalaceae), which occurs at only a single locality in The Netherlands. Stems of this species lay flat on the vegetation, and we have observed that the small, open flowers are visited by ants for nectar. It is not known to what extent ants actually function as pollinators, transferring (cross) pollen from one flower to the next. In addition, T. humifusum has seeds bearing an elaiosome, and we have also observed that ants readily (attempt to) carry them to their nest sites. The main question of this research project is: How important are ants for the pollination and dispersal of this species? Techniques: • Designing and carrying out pollinator exclusion experiments; • Determining the extent of pollen transport by ants using e.g. cubes of sticky gel • Counting fruit and seed set • Performing (cafetaria) experiments to demonstrate (directional) seed dispersal by ants Background reading: Beattie AJ (2006) The evolution of ant pollination systems. Bot. Jahrb. Syst. 127, 43–55. Oostermeijer J.G.B. (1989) Myrmecochory in Polygala vulgaris L., Luzula campestris (L.) DC. and Viola curtisii Forster in a Dutch dune area. Oecologia 78, 302-311. How do genetic processes drive individual fitness in reintroduced populations of endangered plants? Supervisors: Gerard Oostermeijer, Sheila Luijten (Science4Nature), Patrick Meirmans Contact: [email protected] or [email protected] Project outline Many plant and animal populations are threatened with extinction by extensive habitat fragmentation, which reduces population sizes and increases isolation. Inbreeding depression is an important mechanism lowering individual fitness and subsequently population viability. We are trying to alleviate inbreeding depression by genetic rescue, i.e. adding genetic diversity from one or more other (preferably large) populations. When we reintroduce new populations in ecologically restored habitat, we increase genetic diversity by mixing gene pools in a breeding program in the greenhouse or common garden. Although it is quite likely that this reduces inbreeding depression, there is a risk that the mixing of gene pools disrupts local adaptations, resulting in outbreeding depression. We are conducting recovery programs on several endangered plant species, such as Primula vulgaris, Antennaria dioica, Scorzonera humilis, Phyteuma nigrum and Lathyrus linifolius. In the context of these programs, there are many opportunities for Master research projects in which the role of genetic processes like heterosis, inbreeding depression and outbreeding depression can be studied. The detailed backgrounds of each recovery project differ, so that it is best to talk to one of the abovementioned supervisors and discuss a specific research project that best suits your particular interests and time schedule. Techniques: • Plant breeding systems: crossing experiments in the greenhouse and/or in the field • Plant fitness: measuring plant performance traits (fitness components) in the greenhouse and/ or in the field • Genetic diversity: sampling plant leaf tissue in the field or greenhouse and determining genotypes using microsatellites (and in the near future also RADseq/SNPs) in the IBED molecular lab. • Statistical analysis of reproductive success, variation in fitness components and genetic diversity, and relationships between them. Background reading: Barmentlo SH, Meirmans PG, Sheila H. Luijten SH, Triest L, Oostermeijer JGB (2017) Outbreeding depression and breeding system evolution in small, remnant populations of Primula vulgaris: consequences for genetic rescue. Conservation Genetics, in press. Oostermeijer JGB, Luijten SH, den Nijs JCM (2003) Integrating demographic and genetic approaches in plant conservation. Biological Conservation 113, 389-398. Pierson JC, Beissinger SR, Bragg JG, Coates DJ, Oostermeijer JGB, Sunnucks P, Schumaker NH, Trotter MV, Young AG (2015) Incorporating evolutionary processes into population viability models. Conservation Biology 29, 755-764. 2 Ecotypic differentiation between heathland and hay meadow populations of Gentiana pneumonanthe Supervisors: Gerard Oostermeijer, Sheila Luijten (Science4Nature), Patrick Meirmans Contact: [email protected] or [email protected] Project outline Ecotypic differentiation reflects the first stages of speciation, in which populations are adapting to the environmental conditions characterising different habitats. Such differences are very important for conservation biology of endangered species, because mixing ecotypes during genetic rescue, or using the wrong ecotype for reintroduction attempts, will almost certainly decrease the success rate. This is because (a) the genotypes used are less likely to match the environmental conditions and (b) mixing differentially adapted gene pools might result in outbreeding depression by disrupting coadapted (epistatic) gene complexes. In Gentiana pneumonanthe, two putative ecotypes occur, one adapted to wet heathlands and grass heaths and the other to wet hay meadows. In the former, nutrient- poor habitat, demography is driven by episodic recruitment in patches of bare soil created by sod removal, grazing/trampling and/or fire. In this habitat, individual plants need to grow, flower and set seed rapidly, before the vegetation closes by succession. In the more mesotrophic hay meadows, annual mowing and haymaking keeps vegetation succession at bay, and populations are demographically more stable by high survival of established adults, and only occasional recruitment in gaps in the vegetation. The hypothesis is that these two very different types of demographic behaviour are the result of different genetic adaptations. The project entails a comparative study of adaptive traits in heathland and hay meadow populations of G. pneumonanthe in a common garden. In a later stage also a reciprocal transplant experiment will be carried out in the field. Using molecular markers, genetic differentation will also be compared among the studied populations, searching for markers indicating adaptive selection. Techniques: • Plant fitness: measuring plant performance traits (fitness components) in a comparative growth experiment in the greenhouse; • Genetic diversity: sampling plant leaf tissue in the field or greenhouse and determining genotypes using microsatellites (and in the near future also RADseq/SNPs) in the IBED molecular lab. • Statistical analysis of variation in fitness traits and genetic diversity, and relationships between them. Background reading: Oostermeijer JGB, Luijten SH, den Nijs JCM (2003) Integrating demographic and genetic approaches in plant conservation. Biological Conservation 113, 389-398. Hufford KM, Mazer SJ (2003) Plant ecotypes: genetic differentiation in the age of ecological restoration. Trends in Ecology and Evolution 18, 147-155. Pierson JC, Beissinger SR, Bragg JG, Coates DJ, Oostermeijer JGB, Sunnucks P, Schumaker NH, Trotter MV, Young AG (2015) Incorporating evolutionary processes into population viability models. Conservation Biology 29, 755-764. 3 Regeneration niches and the restoration biology of endangered plants Supervisors: Gerard Oostermeijer, Sheila Luijten (Science4Nature), Patrick Meirmans Contact: [email protected] ,or [email protected] Project outline In many cases, recovery projects for endangered plant species involve relatively haphazard reintroduction in what is believed to be good habitat. The definition of good habitat is at best based on old vegetation relevés or descriptions in papers, but rarely includes experimental study of the regeneration niche. The latter is defined as the (micro)habitat in which a species can successfully germinate, establish and grow up to become a flowering adult that can eventually produce new offspring. Management, soil conditions, hydrology, vegetation structure, the timing of sowing (or seed release) and seed viability are all important variables determining a species’ regeneration niche. In many reintroduction projects, conservation biologists do not introduce seeds, but plant out pregrown adult plants. The main drawbacks of this approach are that (a) all genetic-environmental
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